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Accumulation of glutamine synthetase during early development of maritime pine (Pinus pinaster) seedlings

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Abstract

Seedlings of Pinus pinaster Alton accumulated chlorophyll (Chl) when grown in complete darkness. Contents of Chl a and Chl b increased during germination, reaching similar levels in light- and dark-grown plants. Glutamine-synthetase (GS; EC 6.3.1.2) activity was detected in the embryo and its level increased markedly in cotyledons of dark-germinated seedlings. Similar levels of GS activity were observed when the seeds were germinated in the presence of white light. Only one GS form, which eluted at about 0.1 M KCl, was found by ion-exchange chromatography. A predominant GS polypeptide of 43 kDa was detected in cotyledons, and its steady-state level increased with development in a lightindependent fashion. In roots and needles, a related GS polypeptide of 43 kDa was the unique species detectable by western blot analysis. Immunoblots of soluble proteins from isolated chloroplasts showed low abundance of GS protein, indicating that glutamine synthesis in pine cotyledons occurs mainly in the cytosol. Nitrogen-feeding experiments carried out with detached shoots indicated that neither NO −3 nor NH +4 regulate GS levels and the polypeptide pattern. Our results indicate that environmental factors, such as light and nitrogen supply, have a limited role in GS accumulation during pine development.

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Abbreviations

Chl:

Chlorophyll

DEAE:

diethylaminoethyl

GS:

glutamine synthetase

kDa:

kilodalton

Mr :

relative molecular mass

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecyl sulphate

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Authors and Affiliations

  1. Laboratorio de Bioquímica y Biología Molecular Universidad de Málaga, Campus de Teatinos, E-29071, Malaga, Spain

    Francisco M. Cánovas, Francisco R. Cantón, Fernando Gallardo & Angel García-Gutiérrez

  2. Laboratorio de Microbiología, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos, E-29071, Malaga, Spain

    Antonio de Vicente

Authors
  1. Francisco M. Cánovas
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  2. Francisco R. Cantón
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  3. Fernando Gallardo
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  4. Angel García-Gutiérrez
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  5. Antonio de Vicente
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Additional information

We wish to thank Dr. B. Hirel, Laboratoire Métabolisme et Nutrition des Plantes, INRA, Versalles, France, for kindly providing the antibodies to tobacco GS. We are also grateful to Dr. F.M. Sánchez-Jiménez (Bioquímica, Universidad de Málaga, Málaga, Spain) for her comments and criticisms on the manuscript. This research was supported by a Grant from Direccion General de Investigation Científica y Técnica (PB 89-0579).

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Cánovas, F.M., Cantón, F.R., Gallardo, F. et al. Accumulation of glutamine synthetase during early development of maritime pine (Pinus pinaster) seedlings. Planta 185, 372–378 (1991). https://doi.org/10.1007/BF00201059

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  • DOI: https://doi.org/10.1007/BF00201059

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